RESUMEN
OBJECTIVE: To express and purify H5N1 influenza virus (A/Anhui/1/2005) NP in prokaryotic system and to explore the NP-interacting proteins of human bronchial epithelial cells BEAS-2B in vitro. METHODS: The full length H5N1 NP gene fragment was amplified by PCR, inserted into prokaryotic expression vector (pET30a) to generate NP expression plasmid pET30a-NP. After transforming pET30a-NP into E. coli (BL21), the expression of soluble NP protein was induced by IPTG. The expressed NP protein was purified by two steps with metal chelation chromatography and ion exchange chromatography. Then the total proteins of BEAS-2B cells was extracted for screening the components which have protein-protein interaction with purified NP by pull-down and LC-MS/MS methods. RESULTS: The expression of H5N1 NP protein could be induced by IPTG in bacterial system using expression plasmid pET30a-NP. The soluble NP was purified. Twenty proteins were found by pull-down and LC-MS/MS, the further experiments may be needed to prove protein-protein interaction between them. CONCLUSION: The soluble H5N1 NP fusion protein with high purity was obtained and twenty proteins were found which could interact with it by pull-down and LC-MS/MS.
Asunto(s)
Subtipo H5N1 del Virus de la Influenza A/metabolismo , Gripe Humana/metabolismo , Proteínas de Unión al ARN/aislamiento & purificación , Proteínas de Unión al ARN/metabolismo , Proteínas del Núcleo Viral/aislamiento & purificación , Proteínas del Núcleo Viral/metabolismo , Línea Celular , Humanos , Subtipo H5N1 del Virus de la Influenza A/genética , Gripe Humana/virología , Proteínas de la Nucleocápside , Unión Proteica , Proteínas de Unión al ARN/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/aislamiento & purificación , Proteínas Recombinantes/metabolismo , Proteínas del Núcleo Viral/genéticaRESUMEN
OBJECTIVE: To construct adenovirus vector vaccine against H5N1 influenza virus and study on the immunogenicity. METHODS: In this study, we amplified hemagglutinin (HA) gene sequence of H5N1 influenza virus (A/Anhui/1/2005), then constructed an adenovirus vector vaccine (Adv-HA), followed by tests in BALB/c mice for the immunogenicity with the vaccine and immunization strategies. RESULTS: The recombinate Adv-HA vaccine could effectively induce both humoral and cellular immunity against human H5N1 influenza virus. CONCLUSION: The Adv-HA vaccination against H5N1 influenza is a potential strategy and worthy of further investigation.
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Glicoproteínas Hemaglutininas del Virus de la Influenza/inmunología , Subtipo H5N1 del Virus de la Influenza A/inmunología , Vacunas contra la Influenza/inmunología , Gripe Humana/inmunología , Gripe Humana/prevención & control , Adenoviridae/genética , Adenoviridae/inmunología , Animales , Línea Celular , Femenino , Vectores Genéticos/genética , Vectores Genéticos/inmunología , Glicoproteínas Hemaglutininas del Virus de la Influenza/administración & dosificación , Glicoproteínas Hemaglutininas del Virus de la Influenza/genética , Humanos , Subtipo H5N1 del Virus de la Influenza A/genética , Vacunas contra la Influenza/administración & dosificación , Vacunas contra la Influenza/genética , Gripe Humana/virología , Ratones , Ratones Endogámicos BALB C , Distribución Aleatoria , VacunaciónRESUMEN
The ingestion of lactobacilli is of great importance for the probiotic effect of host gut Peyer's patches (PPs) macrophages. The present study is in time focus on the investigation of the factors determining the ingestion of lactobacilli by PPs macrophages. Physicochemical properties of cell surface and adhesive property of nine Lactobacillus strains were examined in the present work. The association of the bacteria with PPs macrophage was checked with macrophage monolayers on coverslips. The influence of lactobacilli on macrophages phagocytic capacity was also investigated with a neutral red uptake assay in vitro. The results show that the macrophages could ingest lactobacilli in a strain dependent manner, and the most ingested strain is L. plantarum Lp6 compared to other tested strains, which displayed strain specific enhancement on the phagocytic activity of PPs macrophages. And there is no correlation between the physicochemical or adhesive properties of the cell surface and the ingestion. The association of L. plantarum Lp6 with PPs macrophage could be decreased by Protease K treatment. Surface proteins of L. plantarum Lp6 could promote the ingestion of fluorescent latex beads by PPs macrophages. In conclusion, the hydrophobicity of the cell surface might not be the key factor determining the association of lactobacilli with PPs macrophages. Cell surface proteins are the media for the binding L. plantarum Lp6 to macrophages.
Asunto(s)
Lactobacillus/inmunología , Macrófagos/microbiología , Ganglios Linfáticos Agregados/inmunología , Ganglios Linfáticos Agregados/microbiología , Fagocitosis , Animales , Adhesión Bacteriana/inmunología , Adhesión Bacteriana/fisiología , Macrófagos/inmunología , Ratones , Probióticos , Especificidad de la EspecieRESUMEN
AIM: To explore some operative techniques to prevent the occurrence of delayed gastric emptying (DGE) after pylorus-preserving pancreaticoduodenectomy (PPPD). METHODS: One hundred and eighty-six patients in a single medical center who accepted PPPD were retrospectively studied. The incidence of DGE was investigated and the influence of some operative techniques on the prevention of DGE was analyzed. RESULTS: During the operative process of PPPD, the methods of detached drainage of pancreatic fluid and bile and gastric fistulization were used. Postoperatively, six patients suffered DGE among the 186 cases; the incidence was 3.23% (6/186). One of them was complicated with intraabdominal infection at the same time, and two with pancreatic leakage. CONCLUSION: Appropriate maneuvers during operation are essential to avoid postoperative DGE in PPPD. The occurrence of DGE is avoidable. It should not be used as an argument to advocate hemigastrectomy in PPPD.